Chromatography column for the separation of surfactants

Abstract

A chromatographic separation column for characterizing surfactant purity is disclosed. The column includes a separation medium having a particulate porous substrate with monofunctional silane with diisopropyl side chain groups and a pendant cyano functional group held within a column. The porous substrate has a pore size of about 50 Å to about 500 Å and an average particle size of about 1.0 μm to about 100 μm. The column has an inner diameter of about 1.0 to 100 mm, e.g., 4.6 mm and a length from about 10 mm to about 250 mm. Methods of facilitating characterization of polysorbate 80 by providing a sample containing polysorbate 80 and one or more reaction products of polysorbate 80 and a chromatographic separation column are disclosed. Methods of characterizing polysorbate 80 by separating polysorbate 80 and one or more reaction products of polysorbate 80 are also disclosed.

What is claimed is: 
     
         1 . A chromatographic separation column for characterizing surfactant purity, comprising,
 a separation medium comprising a particulate porous substrate having a monofunctional silane with diisopropyl side chain groups and a pendant cyano functional group, the porous substrate having a pore size of about 50 Å to about 500 Å and an average particle size of about 1.0 μm to about 100 μm; and   a column constructed and arranged to hold the separation medium, the column having an inner diameter of about 1.0 to 100 mm and a length from about 10 mm to about 250 mm.   
     
     
         2 . The column of  claim 1 , wherein the surfactant to be characterized comprises a non-ionic surfactant. 
     
     
         3 . The column of  claim 2 , wherein the non-ionic surfactant comprises polysorbate-80. 
     
     
         4 . The column of  claim 1 , wherein the porous substrate comprises a hydrated metal oxide, a hydrated metalloid-oxide, or organic polymer. 
     
     
         5 . The column of  claim 4 , wherein the porous substrate comprises silica. 
     
     
         6 . The column of  claim 1 , wherein the porous substrate has a particle size of about 3.5 μm. 
     
     
         7 . The column of  claim 1 , wherein the porous substrate has a pore size of about 300 Å. 
     
     
         8 . The column of  claim 1 , wherein the inner diameter of the column is 2.1 mm. 
     
     
         9 . The column of  claim 1 , wherein the length is about 50 mm to 150 mm. 
     
     
         10 . The column of  claim 1 , wherein the separation medium can resist degradation over a pH range from 1.0 to 8.0. 
     
     
         11 . The column of  claim 1 , wherein the separation medium can be used in a temperature range of about 20 to 80° C. 
     
     
         12 . The column of  claim 1 , wherein the column can be used at an operating pressure up to about 400 bar. 
     
     
         13 . A method of facilitating characterization of polysorbate 80, comprising:
 providing a sample comprising polysorbate 80 and one or more reaction products of polysorbate 80; and   providing a chromatographic separation column, the column comprising,
 a separation medium comprising a particulate porous substrate having a monofunctional silane with diisopropyl side chain groups and a pendant cyano functional group, the porous substrate having a pore size of about 50 Å to about 500 Å and an average particle size of about 1.0 μm to about 100 μm; and 
 a column constructed and arranged to hold the separation medium, the column having an inner diameter of about 1.0 to 100 mm and a length from about 10 mm to about 250 mm. 
   
     
     
         14 . The method of  claim 13 , wherein the porous substrate of the provided chromatographic separation column comprises silica. 
     
     
         15 . The method of  claim 14 , wherein the porous substrate of the provided chromatographic separation column has a particle size of about 3.5 μm. 
     
     
         16 . The method of  claim 15 , wherein the porous substrate of the provided chromatographic separation column has a pore size of about 300 Å. 
     
     
         17 . The method of  claim 16 , wherein the inner diameter of the provided chromatographic separation column method is 2.1 mm. 
     
     
         18 . The method of  claim 17 , wherein the length of the provided chromatographic separation column is about 50 mm to about 150 mm. 
     
     
         19 . The method of  claim 13 , wherein the one or more reaction products of polysorbate 80 includes one or more of long chain fatty acids, short chain fatty acids, polyoxyethylene, polyoxyethylene sorbitan, aldehydes, ketones, formic acid, acetic acid, and peroxides. 
     
     
         20 . The method of any one of  claims 13-19 , further comprising installing the chromatographic separation column into a liquid chromatographic instrument. 
     
     
         21 . The method of  claim 20 , further comprising providing a mobile phase to effectuate separation of the polysorbate 80 and the one or more reaction products of polysorbate 80. 
     
     
         22 . The method of  claim 21 , wherein the mobile phase comprises one or more of ammonium acetate, ammonium formate, ammonia, formic acid, acetic acid, trifluoroacetic acid, methanol, isopropanol, acetonitrile, ethanol, and acetone. 
     
     
         23 . The method of  claim 22 , further comprising providing instructions for a gradient-based separation using the mobile phase. 
     
     
         24 . A chromatographic separation column for the characterization of surfactants, comprising,
 a separation medium comprising a particulate porous substrate having a monofunctional silane with diisopropyl side chain groups and a pendant cyano functional group, the porous substrate having a pore size of about 300 Å and an average particle size of about 3.5 μm; and   a column constructed and arranged to hold the separation medium, the column having an inner diameter of about 2.1 mm and a length of about 50 mm to 150 mm.   
     
     
         25 . The column of  claim 24 , wherein the surfactant to be separated comprises a non-ionic surfactant. 
     
     
         26 . The column of  claim 25 , wherein the non-ionic surfactant comprises polysorbate 80. 
     
     
         27 . A method of characterizing polysorbate 80, comprising:
 preparing a sample comprising polysorbate 80 and one or more reaction products of polysorbate 80;   injecting the sample into a chromatographic separation column, the column comprising:
 a separation medium comprising a particulate porous substrate having a monofunctional silane with diisopropyl side chain groups and a pendant cyano functional group, the porous substrate having a pore size of about 50 Å to about 500 Å and an average particle size of about 1.0 μm to about 100 μm, the column constructed and arranged to hold the separation medium and having an inner diameter of about 1.0 mm to about 100 mm and a length from about 10 mm to about 250 mm; and 
   eluting the polysorbate 80 and the one or more reaction products of polysorbate 80 using a gradient mobile phase such that the polysorbate 80 and the one or more reaction products of polysorbate 80 are resolved with a resolution of at least three.   
     
     
         28 . The method of  claim 27 , wherein the polysorbate 80 and the one or more reaction products of polysorbate 80 are resolved with a resolution of at least five. 
     
     
         29 . The method of  claim 27 , wherein the particulate porous substrate has a pore size of about 300 Å and an average particle size of about 3.5 μm. 
     
     
         30 . The method of  claim 27 , wherein the column has an inner diameter of about 2.1 mm and a length of about 50 mm. 
     
     
         31 . The method of  claim 27 , wherein the gradient mobile phase comprises ammonium acetate and methanol. 
     
     
         32 . The method of any one of  claims 28-31 , wherein the elution of the polysorbate 80 and the one or more reaction products of polysorbate 80 occurs in less than 10 minutes.